1992 Fiscal Year Annual Research Report

蛋白質/DNA複合体の構造

Research Project

Project/Area Number	04254102
Research Institution	Osaka University
Principal Investigator	箱嶋敏雄大阪大学, 薬学部, 助手 (00164773)
Co-Investigator(Kenkyū-buntansha)	広瀬進国立遺伝学研究所, 教授 (90022730) 月原冨武徳島大学, 工学部, 教授 (00032277) 田中勲北海道大学, 理学部, 助教授 (70093052) 白木原康雄兵庫教育大学, 自然系物理, 助教授 (20150287) 郷通子名古屋大学, 理学部, 教授 (70037290)
Keywords	蛋白質 / DNA / 分子認識 / 三次元構造 / 高次構造 / 転写制御 / 特異性
Research Abstract	多くのDNA結合蛋白質(SSB,DNAポリメラーゼβ,PhoB,CamR,OmpR,H-NS)およびDNAとの複合体(PHO4,papl,DNAポリメラーゼβ,c-Myb,HU)の結晶化に成功したが、中でも、大腸菌由来の一本鎖DNA結合蛋白質SSB、ラット由来のDNAポリメラーゼβの31kDa断片、大腸菌由来の転写活性化蛋白質OmpRのDNA結合ドメイン、および"ヘリックス・ループ・ヘリックス"DNA結合モチーフをもつ酵母由来の転写活性化蛋白質PHO4のDNA結合ドメインと17塩基対のDNAオリゴマーとの複合体は3Åから2.5Åの分解能をもつことが判明し、構造解析を進めた。SSB、DNAポリメラーゼβおよびOmpRは、これらの結晶からX線回折データを収集することに成功し、重原子置換結晶のスクリーニングの段階まで到達した。また、PHO4のDNAの複合体結晶は、重原子置換結晶のX線回折データの収集も完了し、電子密度図の作成の階段まで至っている。これらの4つの結晶については、近い将来、三次元構造が解明される地点まで研究を推進することができた。また、シンクロトロン放射光を利用した新しいワイセンベルグカメラから得られるデータの処理の効率化もはかった。更に、"ヘリックス・ターン・ヘリックス"DNA結合モチーフをもつ転写調節蛋白質の進化機構やRNAポリメラーゼII中のバルナーゼ類似ドメインの発見、真核生物の超らせん化蛋白質の発見、および大腸菌RecAおよび酵母Rad51の機能構造の同定などに成功した。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] T.Hakoshima: "Three-dimensional structure of a mutant ribonuclease T_1(Y45W)complexed with non-coggnizable ribonucleotide,2'GMP,and its comparison with specific complex with 2'GMP." <J.>___-<Mol.>___-<Biol>___-.223. 1013-1028 (1992)
[Publications] T.Amisaki: "Molecular dynamics and free energy perturbation calculations on the mutant of tyrosine 45 to tryptophan in ribonuclease t_-." <Chem.>___-<Pharm.>___-<Bull.>___-. 40. 1303-1308 (1992)
[Publications] W.-C.Lam: "Phosphorescence and optically detected magnetic resonance measurements of the 2'AMP and 2'GMP complexes of a mutant ribonuclease T_12(Y45W)in solution:Correlation with X-ray crystal structures." <Biochemistry>___-. 31. 6756-6760 (1992)
[Publications] T.Hakoshima: "Crystallographic characterization of a PHO4/DNA complex." <J.>___-<Mol.>___-<Biol>___-.229. 566-569 (1993)
[Publications] S.Tateishi: "C-terminal truncated <Escherichia>___- <Coli>___- RecA protein,RecA_<5327> has enhanced DNA binding affinities to single-and double-stranded DNAs:Reguratory role of C-terminal region in DNA binding." <J.>___-<Mol.>___-<Biol>___-.223. 115-129 (1992)
[Publications] T.Horii: "Inhibitory effects on N-and C-terminal truncated <Escherichia>___- <coli>___- <recA>___- gene products on functions of wild-type <recA>___- gene." <J.>___-<Mol.>___-<Biol>___-.223. 105-114 (1992)
[Publications] T.Ogawa: "Functional structure of the RecA protein found by chimera analysis." <J.>___-<Mol.>___-<Biol>___-.226. 651-660 (1992)
[Publications] A.Shinohara: "Rad51 protein involved in repair and recombination in <Saccharomyces>___- <cerevisiae>___- is a RecA-like protein." <Cell>___-. 69. 457-470 (1992)
[Publications] T.Ogawa: "Similarity of the yeast Rad51 filament to the bacterial RecA filament." <Science>___-. (1993)
[Publications] M.Go: "Domains and modules of proteins." <J.>___-<Protein>___- <Chem>___-.11. 389-390 (1992)
[Publications] H.Yanagawa: "Protein anatomy:Functional roles in barnase module." <J.>___-<Biol.>___-<Chem>___-.(1993)
[Publications] K.Yoshida: "Protein anatomy:Spontaneous formation of filamentous helical structure from the N-terminal module of barnase." <Biochemistry>___-. (1993)
[Publications] 福山恵一: "ウィルスの立体構造" <バイオサイエンスとインダストリー>___-. 50. 27-30 (1992)
[Publications] S.Yoshikawa: "Strategies for crystallization of large membrane protein complexes." <J.>___-<Crystal>___- <Growth>___-. 122. 298-302 (1992)
[Publications] K.Shinzawa-Itoh: "Single Crystal of bovine heart cytochrome c oxydase at fully oxydized resting,fully reduced and CO-bound fully reduced stages are isomorphous with each other." <J.>___-<Mol.>___-<Biol>___-.228. 987-990 (1992)
[Publications] 月原冨武: "分子置換法による球状ウィルスのX線結晶構造解析" <放射光>___-、. (1993)
[Publications] 月原冨武: "超分子複合体のX線結晶構造解析" <蛋白質核酸酵素>___-. (1993)
[Publications] K.Terada: "Production of antibodies specific for double stranded antigen DNA cloned from immune complexes in plasma of a SLE patient." <Biochem.>___-<Biophys.>___-<Res.>___-<Commuun>___-.183. 797-802 (1992)
[Publications] R.Ohta: "DNA supercoiling facilitates the assembly of transcriptionally active chromation on the adenovirous major late promoter." <Biochem.>___-<Biophys.>___-<Res.>___-<Commuun>___-.186. 963-969 (1992)
[Publications] H.Ueda: "A novel DNA binding motif about the zinc finger domain of insect nuclear hormone receptor FTZ-F1 and mouse ELP." <Mol.>___-<Cell.>___-<Biol>___-.
[Publications] H.Okano: "Regulation of <Drosophila>___- neural development by a putative secreted protein." <Differentation>___-.
[Publications] M.Go: "<The>___- <origin>___- <and>___- <Evolution>___- <of>___- <Prokaryotic>___- <and>___- <Eukaryotic>___- <Cells>___-" World-Scienctific Singapore, (1993)
[Publications] S.Hirose: "<Molecular>___- <Biology>___- <of>___- <DNA>___- <Topoisomerases>___- <and>___- <Its>___- <Application>___- <to>___- <Chemotherapy>___-" CRC press,Florida, (1992)
[Publications] S.Hirose: "<Molecular>___- <Structure>___- <and>___- <Life>___-" CRC Press,Florida, 281 (1992)

1992 Fiscal Year Annual Research Report

蛋白質/DNA複合体の構造

Principal Investigator

箱嶋敏雄大阪大学, 薬学部, 助手 (00164773)

Research Products

[Publications] T.Hakoshima: "Three-dimensional structure of a mutant ribonuclease T_1(Y45W)complexed with non-coggnizable ribonucleotide,2'GMP,and its comparison with specific complex with 2'GMP." <J.>_-<Mol.>_-<Biol>___-.223. 1013-1028 (1992)

[Publications] T.Amisaki: "Molecular dynamics and free energy perturbation calculations on the mutant of tyrosine 45 to tryptophan in ribonuclease t_-." <Chem.>_-<Pharm.>_-<Bull.>___-. 40. 1303-1308 (1992)

[Publications] W.-C.Lam: "Phosphorescence and optically detected magnetic resonance measurements of the 2'AMP and 2'GMP complexes of a mutant ribonuclease T_12(Y45W)in solution:Correlation with X-ray crystal structures." <Biochemistry>___-. 31. 6756-6760 (1992)

[Publications] T.Hakoshima: "Crystallographic characterization of a PHO4/DNA complex." <J.>_-<Mol.>_-<Biol>___-.229. 566-569 (1993)

[Publications] S.Tateishi: "C-terminal truncated <Escherichia>_- <Coli>_- RecA protein,RecA_<5327> has enhanced DNA binding affinities to single-and double-stranded DNAs:Reguratory role of C-terminal region in DNA binding." <J.>_-<Mol.>_-<Biol>___-.223. 115-129 (1992)

[Publications] T.Horii: "Inhibitory effects on N-and C-terminal truncated <Escherichia>_- <coli>_- <recA>_- gene products on functions of wild-type <recA>_- gene." <J.>_-<Mol.>_-<Biol>___-.223. 105-114 (1992)

[Publications] T.Ogawa: "Functional structure of the RecA protein found by chimera analysis." <J.>_-<Mol.>_-<Biol>___-.226. 651-660 (1992)

[Publications] A.Shinohara: "Rad51 protein involved in repair and recombination in <Saccharomyces>_- <cerevisiae>_- is a RecA-like protein." <Cell>___-. 69. 457-470 (1992)

[Publications] T.Ogawa: "Similarity of the yeast Rad51 filament to the bacterial RecA filament." <Science>___-. (1993)

[Publications] M.Go: "Domains and modules of proteins." <J.>_-<Protein>_- <Chem>___-.11. 389-390 (1992)

[Publications] H.Yanagawa: "Protein anatomy:Functional roles in barnase module." <J.>_-<Biol.>_-<Chem>___-.(1993)

[Publications] K.Yoshida: "Protein anatomy:Spontaneous formation of filamentous helical structure from the N-terminal module of barnase." <Biochemistry>___-. (1993)

[Publications] 福山恵一: "ウィルスの立体構造" <バイオサイエンスとインダストリー>___-. 50. 27-30 (1992)

[Publications] S.Yoshikawa: "Strategies for crystallization of large membrane protein complexes." <J.>_-<Crystal>_- <Growth>___-. 122. 298-302 (1992)

[Publications] K.Shinzawa-Itoh: "Single Crystal of bovine heart cytochrome c oxydase at fully oxydized resting,fully reduced and CO-bound fully reduced stages are isomorphous with each other." <J.>_-<Mol.>_-<Biol>___-.228. 987-990 (1992)

[Publications] 月原冨武: "分子置換法による球状ウィルスのX線結晶構造解析" <放射光>___-、. (1993)

[Publications] 月原冨武: "超分子複合体のX線結晶構造解析" <蛋白質核酸酵素>___-. (1993)

[Publications] K.Terada: "Production of antibodies specific for double stranded antigen DNA cloned from immune complexes in plasma of a SLE patient." <Biochem.>_-<Biophys.>_-<Res.>_-<Commuun>_-.183. 797-802 (1992)

[Publications] R.Ohta: "DNA supercoiling facilitates the assembly of transcriptionally active chromation on the adenovirous major late promoter." <Biochem.>_-<Biophys.>_-<Res.>_-<Commuun>_-.186. 963-969 (1992)

[Publications] H.Ueda: "A novel DNA binding motif about the zinc finger domain of insect nuclear hormone receptor FTZ-F1 and mouse ELP." <Mol.>_-<Cell.>_-<Biol>___-.

[Publications] H.Okano: "Regulation of <Drosophila>_- neural development by a putative secreted protein." <Differentation>_-.

[Publications] M.Go: "<The>_- <origin>_- <and>_- <Evolution>_- <of>_- <Prokaryotic>_- <and>_- <Eukaryotic>_- <Cells>___-" World-Scienctific Singapore, (1993)

[Publications] S.Hirose: "<Molecular>_- <Biology>_- <of>_- <DNA>_- <Topoisomerases>_- <and>_- <Its>_- <Application>_- <to>_- <Chemotherapy>_-" CRC press,Florida, (1992)

[Publications] S.Hirose: "<Molecular>_- <Structure>_- <and>_- <Life>_-" CRC Press,Florida, 281 (1992)

1992 Fiscal Year Annual Research Report

蛋白質/DNA複合体の構造

Principal Investigator

箱嶋 敏雄 大阪大学, 薬学部, 助手 (00164773)

Research Products

[Publications] T.Hakoshima: "Three-dimensional structure of a mutant ribonuclease T_1(Y45W)complexed with non-coggnizable ribonucleotide,2'GMP,and its comparison with specific complex with 2'GMP." <J.>___-<Mol.>___-<Biol>___-.223. 1013-1028 (1992)

[Publications] T.Amisaki: "Molecular dynamics and free energy perturbation calculations on the mutant of tyrosine 45 to tryptophan in ribonuclease t_-." <Chem.>___-<Pharm.>___-<Bull.>___-. 40. 1303-1308 (1992)

[Publications] W.-C.Lam: "Phosphorescence and optically detected magnetic resonance measurements of the 2'AMP and 2'GMP complexes of a mutant ribonuclease T_12(Y45W)in solution:Correlation with X-ray crystal structures." <Biochemistry>___-. 31. 6756-6760 (1992)

[Publications] T.Hakoshima: "Crystallographic characterization of a PHO4/DNA complex." <J.>___-<Mol.>___-<Biol>___-.229. 566-569 (1993)

[Publications] S.Tateishi: "C-terminal truncated <Escherichia>___- <Coli>___- RecA protein,RecA_<5327> has enhanced DNA binding affinities to single-and double-stranded DNAs:Reguratory role of C-terminal region in DNA binding." <J.>___-<Mol.>___-<Biol>___-.223. 115-129 (1992)

[Publications] T.Horii: "Inhibitory effects on N-and C-terminal truncated <Escherichia>___- <coli>___- <recA>___- gene products on functions of wild-type <recA>___- gene." <J.>___-<Mol.>___-<Biol>___-.223. 105-114 (1992)

[Publications] T.Ogawa: "Functional structure of the RecA protein found by chimera analysis." <J.>___-<Mol.>___-<Biol>___-.226. 651-660 (1992)

[Publications] A.Shinohara: "Rad51 protein involved in repair and recombination in <Saccharomyces>___- <cerevisiae>___- is a RecA-like protein." <Cell>___-. 69. 457-470 (1992)

[Publications] T.Ogawa: "Similarity of the yeast Rad51 filament to the bacterial RecA filament." <Science>___-. (1993)

[Publications] M.Go: "Domains and modules of proteins." <J.>___-<Protein>___- <Chem>___-.11. 389-390 (1992)

[Publications] H.Yanagawa: "Protein anatomy:Functional roles in barnase module." <J.>___-<Biol.>___-<Chem>___-.(1993)

[Publications] K.Yoshida: "Protein anatomy:Spontaneous formation of filamentous helical structure from the N-terminal module of barnase." <Biochemistry>___-. (1993)

[Publications] 福山 恵一: "ウィルスの立体構造" <バイオサイエンスとインダストリー>___-. 50. 27-30 (1992)

[Publications] S.Yoshikawa: "Strategies for crystallization of large membrane protein complexes." <J.>___-<Crystal>___- <Growth>___-. 122. 298-302 (1992)

[Publications] K.Shinzawa-Itoh: "Single Crystal of bovine heart cytochrome c oxydase at fully oxydized resting,fully reduced and CO-bound fully reduced stages are isomorphous with each other." <J.>___-<Mol.>___-<Biol>___-.228. 987-990 (1992)

[Publications] 月原 冨武: "分子置換法による球状ウィルスのX線結晶構造解析" <放射光>___-、. (1993)

[Publications] 月原 冨武: "超分子複合体のX線結晶構造解析" <蛋白質核酸酵素>___-. (1993)

[Publications] K.Terada: "Production of antibodies specific for double stranded antigen DNA cloned from immune complexes in plasma of a SLE patient." <Biochem.>___-<Biophys.>___-<Res.>___-<Commuun>___-.183. 797-802 (1992)

[Publications] R.Ohta: "DNA supercoiling facilitates the assembly of transcriptionally active chromation on the adenovirous major late promoter." <Biochem.>___-<Biophys.>___-<Res.>___-<Commuun>___-.186. 963-969 (1992)

[Publications] H.Ueda: "A novel DNA binding motif about the zinc finger domain of insect nuclear hormone receptor FTZ-F1 and mouse ELP." <Mol.>___-<Cell.>___-<Biol>___-.

[Publications] H.Okano: "Regulation of <Drosophila>___- neural development by a putative secreted protein." <Differentation>___-.

[Publications] M.Go: "<The>___- <origin>___- <and>___- <Evolution>___- <of>___- <Prokaryotic>___- <and>___- <Eukaryotic>___- <Cells>___-" World-Scienctific Singapore, (1993)

[Publications] S.Hirose: "<Molecular>___- <Biology>___- <of>___- <DNA>___- <Topoisomerases>___- <and>___- <Its>___- <Application>___- <to>___- <Chemotherapy>___-" CRC press,Florida, (1992)

[Publications] S.Hirose: "<Molecular>___- <Structure>___- <and>___- <Life>___-" CRC Press,Florida, 281 (1992)

箱嶋敏雄大阪大学, 薬学部, 助手 (00164773)

[Publications] T.Hakoshima: "Three-dimensional structure of a mutant ribonuclease T_1(Y45W)complexed with non-coggnizable ribonucleotide,2'GMP,and its comparison with specific complex with 2'GMP." <J.>_-<Mol.>_-<Biol>___-.223. 1013-1028 (1992)

[Publications] T.Amisaki: "Molecular dynamics and free energy perturbation calculations on the mutant of tyrosine 45 to tryptophan in ribonuclease t_-." <Chem.>_-<Pharm.>_-<Bull.>___-. 40. 1303-1308 (1992)

[Publications] T.Hakoshima: "Crystallographic characterization of a PHO4/DNA complex." <J.>_-<Mol.>_-<Biol>___-.229. 566-569 (1993)

[Publications] S.Tateishi: "C-terminal truncated <Escherichia>_- <Coli>_- RecA protein,RecA_<5327> has enhanced DNA binding affinities to single-and double-stranded DNAs:Reguratory role of C-terminal region in DNA binding." <J.>_-<Mol.>_-<Biol>___-.223. 115-129 (1992)

[Publications] T.Horii: "Inhibitory effects on N-and C-terminal truncated <Escherichia>_- <coli>_- <recA>_- gene products on functions of wild-type <recA>_- gene." <J.>_-<Mol.>_-<Biol>___-.223. 105-114 (1992)

[Publications] T.Ogawa: "Functional structure of the RecA protein found by chimera analysis." <J.>_-<Mol.>_-<Biol>___-.226. 651-660 (1992)

[Publications] A.Shinohara: "Rad51 protein involved in repair and recombination in <Saccharomyces>_- <cerevisiae>_- is a RecA-like protein." <Cell>___-. 69. 457-470 (1992)

[Publications] M.Go: "Domains and modules of proteins." <J.>_-<Protein>_- <Chem>___-.11. 389-390 (1992)

[Publications] H.Yanagawa: "Protein anatomy:Functional roles in barnase module." <J.>_-<Biol.>_-<Chem>___-.(1993)

[Publications] 福山恵一: "ウィルスの立体構造" <バイオサイエンスとインダストリー>___-. 50. 27-30 (1992)

[Publications] S.Yoshikawa: "Strategies for crystallization of large membrane protein complexes." <J.>_-<Crystal>_- <Growth>___-. 122. 298-302 (1992)

[Publications] K.Shinzawa-Itoh: "Single Crystal of bovine heart cytochrome c oxydase at fully oxydized resting,fully reduced and CO-bound fully reduced stages are isomorphous with each other." <J.>_-<Mol.>_-<Biol>___-.228. 987-990 (1992)

[Publications] 月原冨武: "分子置換法による球状ウィルスのX線結晶構造解析" <放射光>___-、. (1993)

[Publications] 月原冨武: "超分子複合体のX線結晶構造解析" <蛋白質核酸酵素>___-. (1993)

[Publications] K.Terada: "Production of antibodies specific for double stranded antigen DNA cloned from immune complexes in plasma of a SLE patient." <Biochem.>_-<Biophys.>_-<Res.>_-<Commuun>_-.183. 797-802 (1992)

[Publications] R.Ohta: "DNA supercoiling facilitates the assembly of transcriptionally active chromation on the adenovirous major late promoter." <Biochem.>_-<Biophys.>_-<Res.>_-<Commuun>_-.186. 963-969 (1992)

[Publications] H.Ueda: "A novel DNA binding motif about the zinc finger domain of insect nuclear hormone receptor FTZ-F1 and mouse ELP." <Mol.>_-<Cell.>_-<Biol>___-.

[Publications] H.Okano: "Regulation of <Drosophila>_- neural development by a putative secreted protein." <Differentation>_-.

[Publications] M.Go: "<The>_- <origin>_- <and>_- <Evolution>_- <of>_- <Prokaryotic>_- <and>_- <Eukaryotic>_- <Cells>___-" World-Scienctific Singapore, (1993)

[Publications] S.Hirose: "<Molecular>_- <Biology>_- <of>_- <DNA>_- <Topoisomerases>_- <and>_- <Its>_- <Application>_- <to>_- <Chemotherapy>_-" CRC press,Florida, (1992)

[Publications] S.Hirose: "<Molecular>_- <Structure>_- <and>_- <Life>_-" CRC Press,Florida, 281 (1992)